Resident Memory T Cells (TRM) Are Abundant in Human Lung: Diversity, Function, and Antigen Specificity (original) (raw)
Figure 4
Large numbers of immunocompetent and influenza-specific T cells resides in human lung.
Lung T cells were extracted by lung explant method. (A–B) The cytokine secretion of effector memory T cells after overnight stimulation with artificial APCs (microbeads coated with anti-CD2, anti CD3 and antiCD28 mAbs) at 1∶1 cells: bead ratio. Brefeldin A (golgi-stop) was added 6 h prior to intracellular staining of cytokines. A representative dot plot of each cytokine is shown and 6 additional experiments produced similar results. (C) CD4+T cells were stained with TNFα, IL-2 and IFNγ after stimulation with PMA+ionomycin for 6h in presence of brefeldin A (Gate on CD4+IFNγ+ population). A representative dot plot is shown and 10 additional experiments produced similar results. (D) Lung T cells were isolated and stained for different Vbeta T cell receptors using TCR V beta repertoir kit (Beckman coulter) according to manufacturer's instructions. Diversity of V beta TCRs was analyzed by flow cytometry. Data represent Mean +/− SEM of 3 different donors. (E) CFSE labeled T cells from lung, skin and blood were cultured with heat killed influenza virus pulsed APCs in 1∶2 ratio. On day 4, T cell proliferation was measured by analyzing CFSE dilution using flow cytometry. A representative experiment is shown and 2 additional experiments produced similar results.